摘要
目的 :制备人重组P5 3蛋白。方法 :使用含人p5 3基因的重组质粒pT7Hup5 3转化大肠杆菌BL2 1(DE3) ,于培养过程中加入异丙基 β D硫代半乳糖苷诱导转化菌表达目的蛋白P5 3,然后采用溶菌、盐酸胍抽提等方法提取纯化目的蛋白。结果 :最终提取物经蛋白电泳 (SDS PAGE)和免疫印记 (Westernblot)鉴定 ,确为P5 3蛋白 ,含量约0 .373g/L。
Aim:To prepare human recombination P53 protien.Method:Use the recombinant plasmid pT7 7 Hup53 with human p53 cDNA to transform E coli BL21(DE3). During the culture, add into IPTG to induce the transform bacterium to express destination protein P53. And then, with these methods of lysing bacterium and extracted by guanidine hydrochloride extract and purify the destination protein. Result: It was confirmed by protein electrophoresis (SDS PAGE) and immunoblot (Western blot) that the final extract was human P53 protein, and the content was about 0.373 g/L.
出处
《河南医科大学学报》
2000年第3期211-214,F003,共5页
Journal of Henan Medical University
基金
河南省科委自然科学基金资助项目!9840 2 0 3 0 0
